FAQ: What primers should I use to sequence the ends of my insert after I clone it into a pMAL vector?

The following sequencing primers can be used (not available from New England Biolabs): malE primer on the 5′ side of the insert and the pMAL reverse primer for sequencing from the 3′ side.

 Forward Primer (24-mer, upstream of MCS)

 5′d(GGTCGTCAGACTGTCGATGAAGCC)3′

 Reverse Primer (24-mer, downstream of MCS)

 5′d(TGTCCTACTCAGGAGAGCGTTCAC)3′

The sequences of the pMAL vectors are also available at www.neb.com. PCR of inserts cloned into the MCS can be performed using the same primer pair.
Top